Dataset Description

The sample designations correspond to the strains described in the paper as follows: WT represents the parental strain Mc-1; E8 line represents DAOC-1 (∆AccefEF::SccefE/+AccefEF, heterokaryotic mutant); E8 represents DAOC-4 (∆AccefEF::SccefE, homozygous mutant); and ED13 represents DAOC-5 (∆AccefEF::[SccefE-SccefD], homozygous multi-gene-edited mutant).The dataset includes the following contents: (1) PCR validation gel electrophoresis images and corresponding Sanger sequencing data (Tsingke Biotechnology, January/March/June 2025) for mutant strains, using three primer pairs (1-F/R, 2-F(1F)/R, and 0-F/R) to verify gene replacement events; (2) Quantitative HPLC chromatograms (Waters, February/April/July 2025) of DAOC in fermentation broth, using the external standard method for quantification; (3) Qualitative LC-MS spectra (Waters, February/April/July 2025) of fermentation products, identifying compounds based on molecular weight and retention time; (4) qPCR transcriptional data (7500 Fast, July/November 2025), calculated using the 2^（-ΔΔCt） method to determine relative gene expression levels; (5) Whole-genome sequencing data (Meiji Biotechnology, October 2025) for DAOC-4 and DAOC-5, used to assess off-target effects; and (6) Four unpublished data files based on the dual-plasmid system.Data availability notes: The raw process data for the specific DAOC production (mg/g DCW) curves (February/April/July 2025 batches) are unavailable due to failure to back up the data prior to the experimenter's graduation departure.

Additionally, the following qPCR data points are missing: AccefEF expression in Mc-1 at 120 h/144 h, AccefEF expression in DAOC-1 at 120 h/144 h, and SccefE expression in DAOC-1 at 120 h/144 h (July 2025 batch), as these data were deleted due to automatic cleanup of the instrument-associated computer with insufficient storage space, and were not transferred to another storage location within the valid period. It should be emphasized that the missing data are auxiliary and correspond to non-critical time points.